Small-molecule hydrophobic tagging of fusion proteins and induced degradation of same

Patent metadata

Field	Value
Publication number	US-9632089-B2
Application number	US-201615343640-A
Country	US
Kind code	B2
Filing date	Nov 4, 2016
Priority date	Dec 7, 2010
Publication date	Apr 25, 2017
Grant date	Apr 25, 2017

Abstract

The present invention includes compounds that are useful in perturbing or disrupting the function of a transmembrane or intracellular protein, whereby binding of the compounds to the transmembrane or intracellular protein induces proteasomal degradation of the transmembrane or intracellular protein. The present invention further includes a method of inducing proteasomal degradation of a transmembrane or intracellular protein. The present invention further includes a method of identifying or validating a protein of interest as a therapeutic target for treatment of a disease state or condition.

First claim

The invention claimed is:

1. A compound of formula HYD-L R , wherein:
HYD is a hydrophobic group selected from the group consisting of HTL-5, HTL-14 and HTL-18:

and
L R is a linker group having a reactive moiety selected from the group consisting of

and is capable of forming a covalent link between the HYD and a target protein of interest.

2. The compound of claim 1 , wherein the reactive moiety is a haloalkyl group optionally substituted with a monoether or diether group.

3. The compound of claim 1 , wherein the target protein of interest comprises at least one selected from the group consisting of a haloalkane dehalogenase, O6-alkylguanine-DNA alkyltransferase, ACP synthase, SCP synthase, and SFP synthase.

4. The compound of claim 1 , wherein the HYD has a Clog P of at least 4.0.

5. The compound of claim 1 , wherein the target protein of interest is at least one selected from the group consisting of a structural protein, a receptor, an enzyme, a cell surface protein, a behavioral protein, a cell adhesion protein and a protein involved in a biological activity,
wherein the biological activity is at least one selected from the group consisting of catalytic activity, aromatase activity, motor activity, helicase activity, metabolic processes, antioxidant activity, proteolysis, biosynthesis, kinase activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase activity, ligase activity, enzyme regulator activity, signal transducer activity, structural molecule activity, binding activity, cell motility, membrane fusion, cell communication, regulation of biological processes, development, cell differentiation, response to stimulus, cell death, protein transporter activity, nuclear transport, ion transporter activity, channel transporter activity, carrier activity, permease activity, secretion activity, electron transporter activity, pathogenesis, chaperone regulator activity, nucleic acid binding activity, transcription regulator activity, extracellular organization and biogenesis activity and translation regulator activity.

6. A method of determining whether a protein of interest is a target of a bioactive agent or drug, the method comprising:
exposing a cell that utilizes the protein of interest to a protein of interest which is covalently modified with the compound of claim 1 and is present within or on the surface of the cell, whereby the HYD induces degradation of the protein of interest intracellularly or on the surface of cell;
determining if the degradation of the covalently labeled protein modulates the biological activity of the cell through a change in a phenotypic response of the cell, wherein, if the degradation of the covalently labeled protein modulates the biological activity of the cell through a change in a phenotypic response of the cell, the protein of interest is identified as a target for a bioactive agent or drug that treats or prevents a disease and/or condition modulated through the protein of interest.

7. A method of inducing degradation of a protein of interest in a cell, the method comprising reacting, intracellularly or on the surface of the cell, the expressed protein of interest with a compound comprising the compound of claim 1 , wherein the compound of claim 1 forms a covalent bond with the protein of interest, thus yielding a hydrophobically labeled protein, which undergoes degradation.

Assignees

• Univ Yale

Inventors

• Crews Craig M
• Tae Hyun Seop
• Schneekloth Ashley R
• Neklesa Taavi
• Sundberg Thomas

Classifications

• C07K2319/95

  containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence) · CPC title

• C07K1/13

  Labelling of peptides · CPC title

• G01N33/5008

  for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics · CPC title

• C07K2319/20

  containing a tag with affinity for a non-protein ligand · CPC title

• G01N2500/10

  involving cells · CPC title